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原代破骨細(xì)胞前體細(xì)胞轉(zhuǎn)染操作步驟在已經(jīng)發(fā)表文章中的經(jīng)驗(yàn)分享介紹

西部戰(zhàn)區(qū)總醫(yī)院藥學(xué)部，在2021年發(fā)表文章到Cell Communication and Signaling的學(xué)術(shù)期刊中科研者使用美國invigentech轉(zhuǎn)染試劑，高效率轉(zhuǎn)染原代破骨細(xì)胞前體細(xì)胞（Osteoclast precursor cells），具體轉(zhuǎn)染原代破骨細(xì)胞前體細(xì)胞方法經(jīng)驗(yàn)如下：（注意：本文轉(zhuǎn)染實(shí)驗(yàn)中使用的細(xì)胞數(shù)量、試劑用量和操作方法不適用于傳統(tǒng)lipo3000轉(zhuǎn)染試劑或類似產(chǎn)品）。

Cell culture

Sorted primary CD115(+) precursors were cultured in α-minimal essential medium (MEM) containing 10% FBS,1% penicillin-streptomycin solution and 50 ng ml? 1 colony stimulating factor (M-CSF, Abcam). After 3 days,the medium was replaced with fresh medium. LA was added at a concentration of 100 μg/ml to stimulate the precursors. To inhibit the expression of Cadherin-11, the primary osteoclast precursors were transfected with Cadherin-11 siRNA using INVI DNA RNA Transfection reagent (Invigentech) for 24 h. The sequence of Cadherin-11 siRNA was CCAATGGACCAAGATTTAT. In some experiments, primary cells were treated with BYL719 (2.5 μM), GSK690693 (25 nM), 666–15 (50 nM), rapamycin (20 μM), or AMG-487 (30 μM).

Invigentech.英克: 簡介

美國 Invigentech（英克）公司是開發(fā)用于細(xì)胞培養(yǎng)的胎牛血清、細(xì)胞活力檢測、細(xì)胞篩選、細(xì)胞支原體清除、細(xì)胞轉(zhuǎn)染和組織再生的新型血清替代品；提供inviCELL™Platelet lysate無動(dòng)物血清產(chǎn)品用于支持廣泛的細(xì)胞擴(kuò)增和生產(chǎn)，包括培養(yǎng)間充質(zhì)干細(xì)胞和多種免疫細(xì)胞系等，為制藥公司或者生物技術(shù)公司提供無血清細(xì)胞培養(yǎng)規(guī)模生產(chǎn)服務(wù)；INVI DNA  RNA Transfection Reagent為原代細(xì)胞、懸浮細(xì)胞、小動(dòng)物活體轉(zhuǎn)染提供高品質(zhì)轉(zhuǎn)染試劑；此外我們引進(jìn)的新設(shè)施可生產(chǎn)高純度注射用水和生物處理解決方案，用于滿足任何細(xì)胞、科研和企業(yè)客戶需求，每個(gè)批次胎牛血清、細(xì)胞活力檢測、細(xì)胞篩選細(xì)胞，轉(zhuǎn)染等產(chǎn)品質(zhì)量的穩(wěn)定性，以加速安全有效的基于細(xì)胞和組織的治療研究、開發(fā)和商業(yè)化。